Table 1.
Primers used for site-directed mutagenesis of PVX mutants
| Mutant | Primer | Sequences |
|---|---|---|
| V45R | Forward | 5′-CTCAAACGCGTGAAGCGGCTAATGATCTAGAGGGGTTCGGCATAG-3′ |
| Reverse | 5′-GCCGCTTCACGCGTTTGAGCGTACGGGTTAGATAGTTTGTGTTTTTCC-3′ | |
| E46A | Forward | 5′-TCAAACGGTTGCCGCGGCTAATGATCTAGAGGGGTTCGGCATAGC-3′ |
| Reverse | 5′-ATTAGCCGCGGCAACCGTTTGAGCGTACGGGTTAGATAGTTTGTGTTTTTC-3′ | |
| T140A | Forward | 5′-ACAGAGATCGCCACGGACACAGCATACATTAGTGACACTCTGCACTTC-3′ |
| Reverse | 5′-GTGTCCGTGGCGATCTCTGTGAGCTTGTCTATTATTGTTTCCTTTGGG-3′ | |
| D200A | Forward | 5′-TACTTTGGAGCCGGTTTCCAGTATATACCAGGCAACCATGGTGG-3′ |
| Reverse | 5′-TACTGGAAACCGGCTCCAAAGTATTTGAGGCTGTATATGTTCGGGT-3′ | |
| N243A | Forward | 5′-CTCGGACATCTCGCCTACACGACTGAGCAGGTTGAGATGCACACAG-3′ |
| Reverse | 5′-AGTCGTGTAGGCGAGATGTCCGAGAAAGCTATCCTTGGGGTCCCTC-3′ | |
| K299A | Forward | 5′-ATCTTTCTCCCGGCAGTTCACAACTGCAAGAAGCCGATTCTGAAGA-3′ |
| Reverse | 5′-GTTGTGAACTGCCGGGAGAAAGATCTGTGGTGGAATCACATACCTGTCA-3′ | |
| V300R | Forward | 5′-CTCCCGAAACGCCACAACTGCAAGAAGCCGATTCTGAAGAAAAC-3′ |
| Reverse | 5′-CAGTTGTGGCGTTTCGGGAGAAAGATCTGTGGTGGAATCACATACC-3′ | |
| S341R | Forward | 5′-ATTAAATCGCGCGACTTGGACAAATACTCTGCTGTGGAACTGGTTTAC-3′ |
| Reverse | 5′-GTCCAAGTCGCGCGATTTAATTAATTGTCTGACTTTGGCAAAAATG-3′ | |
| D344A | Forward | 5′-GTCTGACTTGGCGAAATACTCTGCTGTGGAACTGGTTTACTTAGTGAGC-3′ |
| Reverse | 5′-GCAGAGTATTTCGCCAAGTCAGACGATTTAATTAATTGTCTGACTTTGGCA-3′ | |
| E446A | Forward | 5′-TCGGACGTAGCCGAAATGGAAAGTTTGTTCTCAGATGGGGACCTG-3′ |
| Reverse | 5′-TTTCCATTTCGGCTACGTCCGACACTTCCCTTGGTCGGAAGG-3′ | |
| A494K | Forward | 5′-ATTAAAGAACCTAAAGGAGACAGAAATCAATACTCAAACCCTGCGGAA-3′ |
| Reverse | 5′-TTCTGTCTCCTTTAGGTTCTTTAATTCCTTGACCGACATCCATCTCA-3′ | |
| K525A | Forward | 5′-CAGACCACAGCGAAGGCTAAGCGCCTAGCTGAAATCCAGGAGTCC-3′ |
| Reverse | 5′-GCTTAGCCTTCGCTGTGGTCTGATGTTTCACCTCTCTACTGTGTTTCCTGT-3′ | |
| K526A | Forward | 5′-CAGACCACAAAGGCGGCTAAGCGCCTAGCTGAAATCCAGGAGTCC-3′ |
| Reverse | 5′-GCTTAGCCGCCTTTGTGGTCTGATGTTTCACCTCTCTACTGTGTTTCCTGT-3′ | |
| K528A | Forward | 5′-AAAGAAGGCTGCGCGCCTAGCTGAAATCCAGGAGTCCATGA-3′ |
| Reverse | 5′-TAGGCGCGCAGCCTTCTTTGTGGTCTGATGTTTCACCTCTCTACTG-3′ | |
| E540A | Forward | 5′-ATGAGAGCAGCCGGTGAAGCTGAATCAAATGAGATGAGCGGG-3′ |
| Reverse | 5′-AGCTTCACCGGCTGCTCTCATGGACTCCTGGATTTCAGCTAGGC-3′ | |
| S545A | Forward | 5′-GAAGCAAATGAGATGAGCGGGGGCATGGGGGCAATACCG-3′ |
| Reverse | 5′-CCCGCTCATCTCATTTGCTTCAGCTTCACCTTCTGCTCTCATGGACT-3′ | |
| S565A | Forward | 5′-AGCACGGCTGATGCTAGACAAGAACTCACACTCCCAACCACC-3′ |
| Reverse | 5′-GTCTAGCATCAGCCGTGCTGGGAAGTTCAGCGTTGCTCG-3′ | |
| D566A | Forward | 5′-AGCACGAGTGCCGCTAGACAAGAACTCACACTCCCAACCACCAA-3′ |
| Reverse | 5′-TCTTGTCTAGCGGCACTCGTGCTGGGAAGTTCAGCGTTGCTCG-3′ | |
| M610K | Forward | 5′-GAGACAGCAAAACAACAAGTCATCGAAGGACTCCCTTGGAAA-3′ |
| Reverse | 5′-ACTTGTTGTTTTGCTGTCTCAACAGCTTCTTTTCCAGGGAGCTT-3′ | |
| K620A | Forward | 5′-ACTCCCTTGGGCACACTGGATTCCTCAACTAAACGCTGTTGGATTC-3′ |
| Reverse | 5′-AATCCAGTGTGCCCAAGGGAGTCCTTCGATGACTTGTTGCATTG-3′ | |
| N641A | Forward | 5′-GGGATAGGGCTGGAACAATGATCATGCCTATCACAGAAATGG-3′ |
| Reverse | 5′-ATTGTTCCAGCCCTATCCCTCTGAATTTCTAGCGCCTTGAATC-3′ | |
| E662A | Forward | 5′-ACTTCCCGGCCGGAACTCCAAAAGAGTTGGCACGAGAATTGCTC-3′ |
| Reverse | 5′-TGGAGTTCCGGCCGGGAAGTCCTCTTTTTCCAACCCGGAGA-3′ | |
| N676A | Forward | 5′-TGCTCGCTATGGCGAGAAGTCCTGCCACCATCCCTTTGGAC-3′ |
| Reverse | 5′-GGACTTCTCGCCATAGCGAGCAATTCTCGTGCCAACTCTTTTGGA-3′ | |
| K773A | Forward | 5′-AGATTGGAGTGCGAAAGTGCCCAACACTGAACCATACATGTTCAAGA-3′ |
| Reverse | 5′-GGCACTTTCGCACTCCAATCTAGCCGCAGTTCATTGGTCG-3′ | |
| N777A | Forward | 5′-AAAGTGCCCGCGACTGAACCATACATGTTCAAGACCTATGAAAAGGCAT-3′ |
| Reverse | 5′-GGTTCAGTCGCGGGCACTTTCTTACTCCAATCTAGCCGCAGTTC-3′ | |
| E779A | Forward | 5′-CCAACACTGCCCCATACATGTTCAAGACCTATGAAAAGGCATTAATTGG-3′ |
| Reverse | 5′-AACATGTATGGGGCAGTGTTGGGCACTTTCTTACTCCAATCTAGCCG-3′ | |
| N863A | Forward | 5′-CGATACTATCTCGCCGCCACACACCGCAACAAGAAAGACCTTGC-3′ |
| Reverse | 5′-TGTGTGGCGGCGAGATAGTATCGGCAGTATTTTGAGAAGTACTCTGTCGC-3′ | |
| N917A | Forward | 5′-ACCGGAAGGGCCGACACGTTCACATACGCTGGATGCCAAG-3′ |
| Reverse | 5′-ACGTGTCGGCCCTTCCGGTGCCCATGTACAGCTTTCTCTTTTCA-3′ | |
| N964A | Forward | 5′-CACTTCGTGGCGACAAGTGCAAACTCTTCGGCCTTCTGGGAA-3′ |
| Reverse | 5′-TGCACTTGTCGCCACGAAGTGAATCCTGTCGGTAGCTCTAGAGAGTGC-3′ | |
| N968A | Forward | 5′-CACAAGTGCAGCGTCTTCGGCCTTCTGGGAAAAGTTAGACA GCACCC-3′ |
| Reverse | 5′-GCCGAAGACGCTGCACTTGTGTTCACGAAGTGAATCCTGTCGGT-3′ | |
| E1001A | Forward | 5′-AGCCGGCAGCCGTAGAGCCAATTCGAGAGCCTGAGCCC-3′ |
| Reverse | 5′-GCTCTACGGCTGCCGGCTCGTACTCCTTGAGTGCTTGTTCTCTC-3′ | |
| R1006A | Forward | 5′-AGCCAATTGCCGAGCCTGAGCCCCAAACACACATGTGTGT-3′ |
| Reverse | 5′-TCAGGCTCGGCAATTGGCTCTACCTCTGCCGGCTCGTACTC-3′ | |
| E1041A | Forward | 5′-ATCCACTCTGCCTCCCATGGCCATTCAAACTGTGTCCAAACTGA-3′ |
| Reverse | 5′-CCATGGGAGGCAGAGTGGATCTCTCTGTCAAACTTTTCCAAAAGTTCC-3′ | |
| S1042A | Forward | 5′-ACTCTGAAGCGCATGGCCATTCAAACTGTGTCCAAACTGAAGACACA-3′ |
| Reverse | 5′-AATGGCCATGCGCTTCAGAGTGGATCTCTCTGTCAAACTTTTCCAAAAG-3′ | |
| K1065A | Forward | 5′-TCAACAAGCAGCGGATGAGACCCTCCTCTGGGCGACCATAGATG-3′ |
| Reverse | 5′-GGTCTCATCCGCTGCTTGTTGATGCGAAAA CAGCTGGATGGTT-3′ | |
| S1092A | Forward | 5′-AATTCTTGGCCAAGAAGGACATTGGAGACGTCCTGTTTCTAAAC-3′ |
| Reverse | 5′-GTCCTTCTTGGCCAAGAATTCTCGGAAGTTTGTTTCTTGATTG-3′ | |
| N1143A | Forward | 5′-AACTTGATTGCGGGGACTGTGAGACAGAGCCCAGACTTTGAT-3′ |
| Reverse | 5′-CACAGTCCCCGCAATCAAGTTGCACTTTGACTTACTGAGGTATTTG-3′ | |
| D1348A | Forward | 5′-ACAGGCTTGAAGCGAAATTACTCCTCAAGTCGAAGCCTGTAATCACG-3′ |
| Reverse | 5′-GGAGTAATTTCGCTTCAAGCCTGTGGAAACTAAGCTTCACTTCTGGAACG-3′ | |
| K1427A | Forward | 5′-TTGTTCGATGAGGCGCAGTGTCAGGCACATACACTCACTTGCAG-3′ |
| Reverse | 5′-ACACTGCGCCTCATCGAACAAGTCATGCAGAGAGTCCTTGTGGT-3′ | |
| L1456K | Forward | 5′-CAGAAACTTTAAGTAACCGTTAAGTTACCTTAGAGATTTGAATAAGATGG-3′ |
| Reverse | 5′-TAACGGTTACTTAAAGTTTCTGAGGCGGGGAAGTGAGACAGTGCCT-3′ |
Mutated nucleotides are shown in bold type